Bag system for the cryopreservation of body fluids

ABSTRACT

The invention relates to a bag system for the cryopreservation of body fluids, that is blood, bone marrow or umbilical cord blood, the parts of the bag system being sterilized, and separable from one another, wherein liquids which permit and/or assist cryopreservation are located in the bag system and/or may be introduced into the bag system in a sterile form. The bag system, at least, comprising: a device for the direct removal of body fluid ( 1 ) from the living body, a region or number of regions for mixing the liquids with the body fluid and/or for storing the mixture of liquids and body fluid, a number of shut-off elements ( 2 ) and connecting lines ( 3 ) and an inlet element ( 4 ) and/or an outlet element ( 5 ). The bag system is characterized in that the parts of the bag system, which are connected in a communicating manner to one another by the connecting lines ( 3 ), form a closed, sterile system before the bag system is used, and the regions for storing the mixture of liquids and the body fluid are hermetically sealable and separable from one another.

BACKGROUND OF THE INVENTION Field of the Invention

The invention relates to a bag system for the cryopreservation of bodyfluids, that is blood, bone marrow or umbilical cord blood, the parts ofthe bag system being sterilized and separable from one another andliquids which permit and/or assist cryopreservation being located in thebag system and/or able to be introduced into the bag system in a sterileform, at least comprising: a device for the direct removal of body fluidfrom the living body, a region or number of regions for mixing theliquids with the body fluid and/or for storing the mixture of liquidsand body fluid (fluid store), a number of shut-off elements andconnecting lines and an inlet element and/or an outlet element. Afurther aspect of the invention relates to a method of preparing bodyfluids, that is blood, bone marrow or umbilical cord blood, forlong-term storage by cryopreservation in a transporting and storingsystem, at least comprising the steps of: introducing the body fluidinto the transporting and storing system, providing or adding liquidswhich permit and/or assist the cryopreservation, and transferring themixture of liquids which permit and/or assist the cryopreservation andthe body fluid into a region for storage in the transporting and storingsystem.

The cryopreservation of body fluids, that is blood, bone marrow orumbilical cord blood, has assumed immense commercial significance as theconstant development in human medicine has progressed.

Umbilical cord blood and its preservation over a number of years, knownas long-term preservation, are crucially significant in connection withpossible later treatment of the child, youngster or adult with meansbased on the body's own substances, for example in the situation of atransplant. This is because this valuable blood is available only at thebirth of the child. After the umbilical cord has been cut, it stillcontains blood which is not required for the child. In the case in whichlong-term preservation is intended, this blood, often an amount of bloodwith a volume of about 50 to 100 ml, is taken up and prepared forcryopreservation and subsequently frozen by known methods. Theaforementioned small amounts of blood removed must expediently be frozenand stored in such a way that a number of samples can be examined andused independently and at different times from one another, possiblyseveral years apart, by the latest methods then available. Withoutknowledge today of the methods which will be applied then,cryopreservation of all constituents of the body fluid removed isconsidered appropriate.

At present, separate systems are used for the preparation and freezing.For instance, the umbilical cord blood is taken up by a system whichoften comprises one or more removal cannulas, one or two containerswhich contain citrate, and the actual blood collecting container, theaforementioned parts of the system being connected in a communicatingmanner to one another by flexible tubes. Such a system is known as priorart, for example from U.S. Pat. No. 5,879,318.

Following the preparation of the blood, decanting then takes place fromone system into another for the purpose of cryopreservation, that isfrom a sterile container into a sterile freezing container, which oftenhas to be performed under ultraclean room conditions. Assurance of thestatutory predetermined ultraclean room conditions with respect to humanmedicine, for example the requirements according to GMP, often requireshigh investment expenditure, which usually cannot be provided in everydelivery ward or medical facility removing the body fluids in sterileform and preparing them for cryopreservation.

Until now, the preparation for cryopreservation of umbilical cord bloodalways took place by the following steps:

Removal of the blood and introduction into the transporting bag.Addition of citrate, unless already provided, closing of the bag andtransport at room temperature to an ultraclean room within about 24 to48 hours. Transfer of the blood and added cryoprotectant from thetransporting bag into a sterile freezing bag under ultraclean roomconditions, at least two separate systems have always been used.

SUMMARY of THE INVENTION

An object of the invention is to provide a bag system which reduces thecosts of preparing for and carrying out cryopreservation and allows easyand safe manual handling, while having to ensure that the requirementsfor sterility in human medicine are met. Furthermore, it is an object ofthe invention to provide a method of preparing body fluids, that isblood, bone marrow and umbilical cord blood, for long-term storage bycryopreservation which uses a transporting and storing system of theaforementioned type.

The object of the invention is achieved by the parts of the bag systemwhich are connected in a communicating manner to one another by theconnecting lines forming a closed, sterile system before the bag systemis used and the regions for storing the mixture of liquids and the bodyfluid being hermetically sealable and separable from one another.

The invention makes it possible to provide a bag system which forms whatis known as a closed system, at least at the time at which the bodyfluid is introduced, in particular directly from the human body.Consequently, optimum sterility is achieved and there is the possibilityof separating parts of the bag system which are no longer required, inparticular in connection with the actual cryopreservation, step by stepfrom the bag system.

The parts which are no longer required for expedient handling, such asfor example the device for taking up the body fluid after transferringthe body fluid into a region for mixing, are separated from the bagsystem. At the time of freezing, the closed system then only comprisesparts which are separated and hermetically sealed from one another andare no longer connected in a communicating manner to one another.Savings with respect to transporting and storage costs are therebyachieved.

The feature according to the invention that the regions for storing themixture of liquids which permit and/or assist the cryopreservation andthe transferred body fluid are hermetically sealable and separable fromone another makes it possible to separate the total amount of theaforementioned mixture into a number of part-amounts which are separatefrom one another and desired with respect to the size of their volume.These separated part-amounts are available in particular after freezingfor bacteriological and serological examinations and for monitoring thelong-term storage, without opening the complete bag system. The entirehandling in the laboratory, for example the adding of a cryoprotectantvia a sterile filter, can be performed under normal hygienic conditions(for example clean room class D); ultraclean room conditions are notrequired.

The materials used for the parts of the bag system are selected andadapted to one another in a known manner, in particular in respect ofthe function of the respective part, the type of the medium or mediawith which it comes into contact, the acting time of the medium, thetype and manner of the sterilization and the temperature regime, i.e. inparticular the maximum and minimum temperature to be encountered duringhandling and freezing. Known plastics which meet the aforementionedrequirements and are authorized for applications in human medicine aremainly used for this.

Liquids which permit and/or assist the cryopreservation are, for thepurposes of the invention, media known in this respect. For thecryopreservation of blood, they are, for example, liquid media for theprevention of blood coagulation, such as citrate phosphate dextrose(CPD), for the thinning of the blood, such as sodium chloride solutions,and cryoprotectants, such as DMSO.

Regions for mixing the liquids with the body fluid and/or for storageare, for the purposes of the invention, all known closable liquidstores, such as for example containers and bags of medical andlaboratory technology which are formed in such a way as to beliquid-tight with respect to the atmosphere.

In a preferred configuration according to the invention, the regions forstoring the mixture of liquids and the body fluid are configured in sucha way. that they are at least two chambers of a bag. Two chambers permitthe separation of two part-amounts of the liquid mixture to be stored,so that a subdivision which is also expedient for practical purposeswith respect to the complete amount of liquid is possible. The use of abag which has flexible walls is particularly advantageous if thehandling is carried out manually by midwives or medical personnel.Exposure of the flexible walls to manual pressure and the use of gravityhave the effect of transporting the liquid and gas within the bag systemand of bringing about thorough internal mixing.

A further advantageous configuration is that a region for storing themixture of liquids and the body fluid is a connecting line, whichpreferably has a number of segments which can be hermetically sealed andseparated from one another. This configuration achieves the effect thatsmaller part-amounts, which serve in particular as samples formonitoring long-term preservation, are separated in a simple manner, forexample by hermetic sealing by means of welding the flexible andtransparent connecting line which is arranged between the freezing bagand the mixing bag or between the freezing bag and the sterile filterused for venting.

It is also advantageous that a filling level indication is arranged atleast in the respective region for mixing the liquids. The filling levelindication may take place by applying corresponding markings on the wallof the transparent bag, so that in particular the minimum filling heightfor the body fluid introduced and the set filling height, which is to beachieved after filling with the thinning liquid, can be monitored.

In connection with long-term preservation, it is also advantageous thatthe regions for storing the mixture of liquids and the body fluid whichhave been separated from the other parts of the system can be surroundedby an enclosure 8. This enclosure serves in particular for mechanicalprotection and may additionally ensure gas-tight and liquid-tightenclosure.

It is particularly advantageous that, in particular with regard to itsdimensioning and material selection, the bag system is intended forbeing used once. Disposable systems or their parts have a range of knownadvantages, knowledge that they will be used only once, possibly usedonly briefly, such as for example the device for removing the bodyfluid, permitting inexpensive manufacture of the part or system.

For certain applications it is advantageous that the region for mixingthe liquids with the body fluid and the region for storing the mixtureof liquids and body fluid are arranged in a mixing and freezing bag 16,the mixing and freezing bag 16 having a mixing and freezing chamber 9 ora mixing and freezing chamber 9 and a freezing chamber 10.

Fewer parts of the bag system are less expensive and permit easierhandling. This is the case for example whenever it is ensured thatfreezing begins within a relatively short time, i.e. within 10 to 20minutes, after the body fluid has been taken up into the bag system.

Alternatively, the following configuration of the bag system accordingto the invention is preferred, that is that the region for mixing theliquids with the body fluid is arranged in a mixing bag 11 and theregion for storing the mixture of liquids and body fluid is arranged ina freezing bag 12, this bag having a freezing chamber 9 or two freezingchambers 9.

The use of a number of bags, that is one for mixing and one for storing,which may consist of different materials, brings about better handlingand consequently enhanced functional dependability by eliminatingsubjective manipulation errors. For example, specific material selectionmakes it possible in the case of a bag system for umbilical cord bloodto provide CPD in a mixing bag already while the complete bag system isbeing assembled, so that there is no longer any need for otherwisesterile filling of CPD into the bag system immediately before theumbilical cord blood is taken up in the delivery room. It is alsoconsequently possible that the freezing must be performed only withinabout 20 hours after the take-up of the blood into the bag system ifsignificant damage to constituents of the blood is not to occur,allowing intermediate storage at room temperature before freezing. Theselection of the material for this freezing bag 12 is primarilydetermined by its temperature resistance with respect to the actualcryopreservation (temperatures down to about −196° C.); sterilization byautoclaving is consequently often ruled out.

For the aforementioned form of the bag system with two bags for mixingand storing, it is further preferred that the mixing bag 11 and thefreezing bag 12 are connected to each other by two connecting lines.Blood or umbilical cord blood has a tendency to foam, in particularduring mixing with other liquids, i.e. air bubbles are trapped in theliquid or in the liquid mixture, with the result that a liquid-airmixture is obtained. With this type of arrangement, i.e. a closedcircuit between the two bags, the liquid-air mixture contained in thebag system can be manipulated in such a way that there are no or fewinclusions of air in the liquid at the time of freezing in the amount ofliquid which is located in the region or the regions for storing.

Alternatively, it is advantageous that the mixing bag 11 and thefreezing bag 12 are connected to each other by a connecting line. Thisconfiguration expediently requires the arrangement of an outlet elementon the freezing bag, for example a sterile filter, which serves inparticular for the discharge of air from the bag system.

The object of the invention is also achieved by a method in which theparts of a transporting and storing system which are connected in aliquid-communicating manner to one another form a closed, sterile systembefore the body fluid is introduced into the transporting and storingsystem, and the region or the regions for storing the mixture of liquidsand the body fluid are hermetically sealable and separable from oneanother.

The method according to the invention may alternatively be used forhuman body fluids: blood, bone marrow or umbilical cord blood. Use ofthe method is also suitable in principle for other human or animal bodyfluids.

Use of the method according to the invention is particularlyadvantageous in the case where the transporting and storing system is abag system according to claims 1 to 11. In principle, the method canalso be applied if, instead of bags, the transporting and storing systemuses other known liquid stores of medical and laboratory technology,such as bottles or containers. In this case, it is expedient to usecustomary means, such as pumps, at least for part of the liquidcommunication in the transporting and storing container.

It is particularly economical in terms of the method that a definedamount of the mixture of liquids which permit and/or assist thecryopreservation and the body fluid is transferred into a region forstorage. The transferring and storing of a specific amount of liquid,for example 160 ml, permits easy and consequently low-cost freezing withconventional equipment for cryopreservation.

It is also preferred that the mixture of liquids which permit and/orassist the cryopreservation and the body fluid is transferred virtuallywithout any bubbles into a region for storing. Freedom from bubbles isan important criterion for the quality and, consequently, thesuitability for the intended use of the liquid mixture to be stored.This requires for example the arrangement of an outlet element and/or ofthe corresponding connecting lines, so that manual manipulation can beperformed until the liquid mixture is virtually free from bubbles.

The invention is to be explained in more detail below on the basis of anexemplary embodiment of a bag system for umbilical cord blood. In thedrawing:

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows a bag system with a mixing and freezing bag,

FIG. 2 shows a bag system with a mixing and freezing bag which has amixing and freezing chamber and a freezing chamber,

FIG. 3 shows a bag system with a mixing and freezing bag which has amixing and freezing chamber and a sodium chloride bag,

FIG. 4 shows a bag system with a mixing and freezing bag which has a bagfor a liquid for avoiding blood coagulation and a sodium chloride bag,

FIG. 5 shows a bag system which has a freezing bag with two chambers anda mixing bag and two connecting lines between the freezing bag and themixing bag,

FIG. 6 shows a bag system which has a freezing bag with two chambers anda mixing bag and a connecting line between the freezing bag and themixing bag,

FIG. 7 shows a bag system which has a freezing bag with a chamber, amixing bag and two connecting lines between the freezing bag and themixing bag,

FIG. 8 shows a bag system with a glass ampoule in the protective bag and

FIG. 9 shows parts of a bag system before freezing.

DESCRIPTION OF THE PREFERRED EMBODIMENTS

FIG. 1 shows a bag system with a mixing and freezing bag 16, which isconnected in a communicating manner by three connecting lines 3.1, 3.2and 3.3 to a device for the direct removal of umbilical cord blood 1, aninlet element 4 and an outlet element 5. The communication, i.e. inparticular the liquid transport and the air exchange, between theaforementioned parts of the bag system is realized by opening or closingof the shutoff elements 2, which take the form of commercially availablehose clips. The transport of the liquid media, possibly with airinclusions, is realized in the bag system by using gravity and byexertion of manual pressure on the flexible walls of the bags. The inletelement 4, which in this configuration is a sterile filter, serves forthe sterile introduction of the liquids which permit and/or assistcryopreservation, i.e. in particular a liquid for avoiding bloodcoagulation (A), a liquid for thinning (B) and/or a liquidcryoprotectant (C).

After the introduction of the umbilical cord blood and/or liquids whichpermit and/or assist cryopreservation, the connecting lines 3.1 and 3.2,which consist of a plasticized PVC material, can be hermetically sealedand/or separated by welding, for example with a transportable filmwelding appliance. Like the connecting lines 3, the flexible mixing andfreezing bag 16 substantially consists of an EVA plastics material,ethylene vinyl acetate, which is temperature-resistant at least down toa temperature of −196° C. and can be sterilized by radiationsterilization (gamma radiation). On the transparent freezing bag 16,which is designed for a maximum volume of 180 ml, there is a fillinglevel indication 7, which indicates the values for the minimum fillingamount of 60 ml and the set filling amount of 160 ml. Arranged on themixing and freezing bag 16 is a removal adaptor 13, which permits thesterile removal of the stored liquid mixture. The connecting line 3.3 isconnected to an outlet element 5, which is a sterile filter and servesin particular for the air discharge from the bag system. Thethin-walled, flexible and transparent connecting line 3.3 is a regionfor storing the mixture of liquids and the body fluid. These segments 6,for example five of them, with a capacity of in each case about 1 ml,which are hermetically sealable and separable from one another, notrepresented in FIG. 1, can be produced by welding, for example with atransportable film welding appliance, from part of the connecting line3.3. The joining together of the previously sterilized parts of the bagsystem, which comprise commercially available parts and components, toform a closed system for the purposes of the invention is performedunder ultraclean room conditions. Welding, adhesive bonding and joiningtechniques authorized for human medicine can be used for the joiningtogether.

The bag system represented in FIG. 2 shows a mixing and freezing bag 16,which has a mixing and freezing chamber 9 and a freezing chamber 10,which are connected in a communicating manner to each other. The mixingand freezing bag 16 communicates via a number of connecting lines 3 witha device for the direct removal of umbilical cord blood 1, an inletelement 4 and two outlet elements 5. The two chambers, which can beseparated from each other, that is the mixing and freezing chamber 9 andthe freezing chamber 10, in each case have a removal adaptor 13 and anoutlet element 5.

The configuration presented in FIG. 3 differs from FIG. 1 in that a bag14, which contains 21 ml of CPD, a liquid for avoiding blood coagulation(A), and a bag 15, which contains 100 ml of a sodium chloride solution(NaCl), which are respectively connected to the closed bag system via aconnecting line 3, are arranged. The bags 14 and 15 have in each case aninterruption valve with respect to the connecting line 3.

In FIG. 4, the bag system has a mixing and freezing bag 16, which as adifference from FIG. 3 contains two chambers, that is a mixing andfreezing chamber 9 and a freezing chamber 10.

Represented in FIG. 5 is a bag system comprising a mixing bag 11, whichis connected in a communicating manner by four connecting lines 3.1,3.2, 3.3 and 3.4 to a device for the direct removal of umbilical cordblood 1, an inlet element 4, a freezing bag 12 and a bag 15. Thecommunication, i.e., in particular the liquid transport and the airexchange, between the aforementioned parts of the bag system is realizedby opening or closing the shut-off elements 2. The liquid media,possibly with air inclusions, are mixed, moved and transported in thebag system by using gravity and by exertion of manual pressure on theflexible walls of the bags. The inlet element 4, which in thisconfiguration is a sterile filter, serves for the sterile introductionof a liquid cryoprotectant C, for example DMSO. After the introductionof the umbilical cord blood and/or liquids which permit and/or assistcryopreservation, the connecting lines 3.1 and 3.2 can be hermeticallysealed and/or separated by welding. CPD may be provided in the mixingbag 11. Like the connecting lines 3, the flexible mixing bag 11substantially consists of a plasticized PVC plastics material, which canbe sterilized in a way known, e.g. by autoclaving. On the transparentmixing bag 11, which is designed for a maximum volume of 180 ml, thereis a filling level indication 7, which indicates the values for theminimum filling amount of 60 ml and the set filling amount of 160 ml.Arranged on the freezing bag 12, which substantially consists of an EVAplastics material and has two freezing chambers 10, at each freezingcontainer 10 is a removal adaptor 13, which permits the sterile removalof the stored liquid mixture. The connecting lines 3.3 and 3.4 connectthe mixing bag 11 and the freezing bag 12 to each other. One of thethin-walled, flexible and transparent connecting lines 3.3 or 3.4 may bea region for storing the mixture of liquids and the body fluid. Thesesegments 6, for example five of them, with a capacity of in each caseabout 1 ml, which are hermetically sealable and separable from oneanother, not represented in FIG. 5, can be produced by welding, forexample with a transparent film welding appliance, from part of theconnecting line 3.3 or 3.4. The two freezing chambers 10 are connectedto each other in a communicating manner by a connecting line 3.5. Afterthe hermetic sealing and separating of the connecting lines 3.3, 3.4 and3.5 from the mixing bag 11 and freezing bag 12, the two freezingchambers 10 are separable from each other.

The joining together of the previously sterilized parts of the bagsystem, which comprise commercially available parts and components, toform a closed system for the purposes of the invention is performedunder ultraclean room conditions. Welding, adhesive bonding and joiningtechniques authorized for human medicine can be used for the joiningtogether.

FIG. 6 shows a bag system analogous to FIG. 5, the mixing bag 11 and thefreezing bag 12 being connected by a connecting line 3.4. The absentconnecting line 3.3 is substituted by an outlet element 5, which is asterile filter, which serves in particular for venting.

In FIG. 7 the bag system is represented analogously to FIG. 5, thefreezing bag only having one freezing chamber 10.

Represented in FIG. 8 is a bag system comprising a mixing bag 11, whichis connected in a communicating manner by four connecting lines 3.1,3.2, 3.3 and 3.4 to a device for the direct removal of umbilical cordblood 1, a freezing bag 12 and two bags 15.1 and 15.2. Thecommunication, i.e., in particular the liquid transport and the airexchange, between the aforementioned parts of the bag system is realizedby opening or closing the shut-off elements 2. The liquid media,possibly with air inclusions, are transported in the bag system by usinggravity and by exertion of manual pressure on the flexible walls of thebags. The inlet element 4, which in this configuration is a tapping pinfor a pierceable stopper of a glass ampoule 18, serves for the sterileintroduction of a liquid cryoprotectant C, for example, 10 ml of DMSO.The glass ampoule 18 is arranged in a hermetically sealed protective bag17, which is connected to the bag 15.1 via a connecting line 3.6. Thebag 15.1, which has a capacity of about 20 ml, contains an amount ofabout 10 ml of a sodium chloride solution. With respect to theconnecting lines 3.2 and 3.6, the bag 15.1 is in each case closed by aninterruption valve. The bag 15.2, which is connected by the connectingline 3.2 to the bag 15.1 and the mixing bag 11 and is closed by aninterruption valve, contains 100 ml of a sodium chloride solution. Afterthe introduction of the umbilical cord blood and/or liquids which permitand/or assist cryopreservation, the connecting lines 3.1 and 3.2 can behermetically sealed and/or separated by welding. Like the connectinglines 3, the flexible mixing bag 11 substantially consists of aplasticized PVC plastics material, which can be sterilized in a wayknown, e.g., by autoclaving. The transparent mixing bag 11 is designedfor a maximum volume of 180 ml. Arranged on the freezing bag 12, whichsubstantially consists of an EVA plastics material and has two freezingchambers 10, at each freezing container 10 is a removal adaptor 13,which permits the sterile removal of the stored liquid mixture. Theconnecting lines 3.3 and 3.4 connect the mixing bag 11 and the freezingbag 12 to each other. One of the thin-walled, flexible and transparentconnecting lines 3.3 or 3.4 may be a region for storing the mixture ofliquids and body fluid. These segments 6, for example five of them, witha capacity of in each case about 1 ml, which are hermetically sealableand separable from one another, not represented in FIG. 8, can beproduced by welding from part of the connecting line 3.3 or 3.4.

The two freezing chambers 10 are connected to each other in acommunicating manner by a connecting line 3.5. After the hermeticsealing and separating of the connecting lines 3.3, 3.4 and 3.5 from themixing bag 11 and freezing bag 12, the two freezing chambers 10 areseparable from each other.

FIG. 9 shows parts of a bag system before freezing. After the separationof the hermetically sealed parts of a bag system which are to be passedon for cryopreservation, that is one or two chambers of the freezing bag12 or of the mixing and freezing bag 16 and/or the segments 6, whichcontain a liquid mixture D, comprising umbilical cord blood and liquidswhich permit and/or assist cryopreservation, the introduction of theaforementioned parts of the bag system into an enclosure 8 takes place.This enclosure 8, which serves in particular for protection frommechanical damage, comprises a cold-resistant plastics film which issealed in a gas-tight manner by welding.

LIST OF DESIGNATIONS

-   device for the removal of body fluid (1)-   shut-off elements (2)-   connecting lines (3)-   inlet element (4)-   outlet element (5)-   segments (6)-   filling level indication (7)-   enclosure (8)-   mixing and freezing chamber (9)-   freezing chamber (10)-   mixing bag (11)-   freezing bag (12)-   removal adaptor (13)-   bag with A (14)-   bag with B (15)-   mixing and freezing bag (16)-   protective bag (17)-   glass ampoule (18)-   liquid for avoiding blood coagulation (A)-   liquid for thinning (B)-   cryoprotectant (C)-   liquid mixture (D)

1. A bag system for collection and cryopreservation of a quantity ofbody fluid and cells contained therein in a sterile, closed systemhaving a plurality of parts, the system comprising: pre-connected devicefor directly and sterilely collecting body fluid and cells containedtherein from a subject's body or blood vessel into the previouslysterilized, closed system having a plurality of interconnected sterileparts, separably connected to one another within the closed system,capable of holding or receiving cryopreservation liquids in sterile formwithout damage to the system, wherein the plurality of parts compriseone or more regions; component for holding concentrated cryopreservationliquids in a protected area within the sterile closed system untiladdition of the cryopreservation liquid to the collected body fluidwithin the system; region for mixing the cryopreservation liquids and avolume of the collected body fluid and cells contained therein to form amixture, the region having a closable vent for venting air therefromwithin the closed system without opening the closed system, wherein theregion is airtight when the vent is closed; a plurality of shut-offelements and connecting lines, wherein the connecting linescommunicatingly connect parts of the bag system to one another withinthe closed, sterile system, wherein the regions for storing the liquidsand the body fluid and mixtures thereof are hermetically sealable andseparable from one another without breaching the closed system, andwherein the collected fluids and cells within the sterile, closed bagsystem are not exposed to outside contamination at any time duringcollection, processing or cryopreservation.
 2. The bag system accordingto claim 1, wherein the regions for storing the mixture of liquids andthe body fluid and cells contained therein comprise at least twochambers of a freezing bag.
 3. The bag system according to claim 2,comprising a mixing bag within which is the region for mixing theliquids with the body fluid and cells contained therein, and thefreezing bag within which is a region for storing the mixture of liquidsand body fluid.
 4. The bag system according to claim 3, wherein thefreezing bag comprises one freezing chamber.
 5. The bag system accordingto claim 3, wherein the freezing bag comprises at least two freezingchambers.
 6. The bag system according to claim 3, comprising aconnection between the mixing bag and the freezing bag by two or moreconnecting lines, at least one of which carries the mixture of liquidsand fluids from the mixing bag to the freezing bag or chamber of thefreezing bag, and a second line carries air from the mixture of liquidsand fluids in the freezing bag or chamber, back to the emptied mixingbag.
 7. The bag system according to claim 3, comprising a connectionbetween the mixing bag and the freezing bag by one connecting line. 8.The bag system according to claim 3, configured for cryopreservation ofblood, bone marrow, or umbilical cord blood.
 9. The bag system accordingto claim 1, wherein the regions for storing the mixture of liquids andthe body fluid and cells contained therein comprise at least twochambers of a mixing/freezing bag.
 10. The bag system according to claim1, wherein one or more regions for storing the mixture of liquids andbody fluid and cells contained therein are formed in at least one of theconnecting lines having a plurality of hermetically sealable andseparable segments.
 11. The bag system according to claim 1, furthercomprising a fill level indicator affixed to the region for mixing theliquids.
 12. The bag system according to claim 1, wherein only theregions that are hermetically sealable and separable from one anotherfor storing the mixture of liquids and body fluid and cells containedtherein are configured for storing by cryopreservation.
 13. The bagsystem according to claim 1, wherein the parts of the bag system areconfigured for only single-use.
 14. The bag system according to claim 1,wherein the region for mixing the liquids with the body fluid and theregion for storing the mixture of liquids and body fluid and cellscontained therein comprise a mixing/freezing bag, and wherein themixing/freezing bag further comprises a mixing/freezing chamber.
 15. Thebag system according to claim 1, wherein the region for mixing theliquids with the body fluid and the region for storing the mixture ofliquids and body fluid and cells contained therein comprise amixing/freezing bag, and wherein the mixing/freezing bag furthercomprises a mixing/freezing chamber and a freezing chamber.
 16. A methodof preparing body fluids and cells contained therein for long-termstorage by cryopreservation in a closed, sterile transportation andstorage system, the method comprising: providing a steriletransportation and storage system having parts pre-connected to oneanother in a liquid-communicating manner to form a the closed, sterilesystem; introducing body fluid and cells contained therein into thesterile transportation and storage system; holding concentratedcryopreservation liquids until needed in a protected area within theclosed system where it will not damage the system; mixing variablevolumes of the body fluid with cryopreservation liquids to form amixture within the closed system, and venting air therefrom within theclosed system as necessary to minimize bubbles while maintainingsterility of the mixture; transferring the mixture of cryopreservationliquids and body fluid into a region for storage within the closedtransportation and storage system; and hermetically sealing andseparating the region or the regions for storing the mixture ofcryopreservation liquids and body fluid from the remainder of the closedsystem.
 17. The method according to claim 16, further comprisingprocessing body fluids and cells contained therein selected from thegroup consisting of blood, bone marrow, and umbilical cord blood. 18.The method according to claim 16, wherein the providing step comprisesproviding the transportation and storage system comprising a bag systemfor the cryopreservation of a quantity of body fluid and cells containedtherein, wherein the system has a plurality of sterilized parts,separably connected to one another within the closed system, and capableof holding or receiving cryopreservation liquids in sterile form,wherein the plurality of parts comprise one or more regions within theclosed system, and wherein the system comprises: sterile filtrationmeans for directly removing body fluid and cells contained therein froma subject's body or blood vessel; a plurality of shut-off elements andconnecting lines within the closed system, wherein the connecting linescommunicatingly connect parts of the bag system to one another withinthe closed, sterile system, wherein the regions for storing the liquidsand the body fluid and mixtures thereof are hermetically sealable andseparable from one another without breaching the closed system.
 19. Themethod according to claim 16, further comprising storing a total amountof liquid in the transportation and storage system in a number ofpartial amounts, hermetically separated from one another.
 20. The methodaccording to claim 16, further comprising carrying out all of the methodsteps as manual steps.
 21. The method according to claim 16, furthercomprising transferring a defined amount of the mixture ofcryopreservation liquids and body fluid into a region for storing. 22.The method of claim 16, further comprising: venting air from the mixtureof liquids and fluids in the storage region via a connecting line; andthen proceeding with the step of sealing and separating the line fromthe liquid/fluid filled storage region(s), thereby preventing reentry ofthe air into the storage region(s).
 23. The method of claim 22, furthercomprising: returning air removed from the mixture of liquids and fluidsin the storage region via a connecting line to an emptied region of thesystem and then proceeding with the step of sealing and separating theline from the liquid/fluid filled storage region(s), thereby preventingreentry of the air into the storage region(s).
 24. The method of claim16, further comprising: delivering a portion of the mixture of liquidsand body fluids and cells contained therein into a connecting line ofthe closed system; then hermetically sealing and separating segments ofthe line, thereby dividing the mixture contained therein in aliquots ≦1ml; and then cryopreserving and storing the segmented line togetherwith, or separately from, the remainder of the liquid/fluid mixture,such that a single segment can be separated and the aliquotindependently thawed without disturbing the remainder of thecryopreserved mixture.